Structural analysis of parathormone fragments elaborated by cells cultured from a hyperplastic human parathyroid...

Parathyroid cells were obtained by collagenase digestion of 2 g of human parathyroid tissue obtained at surgery from a patient with end stage renal failure and hypercalcemia. Cells were placed into monolayer culture in supplemented Waymouth's MB752/1. Secretion of parathyroid hormone (PTH) from monolayer cultures was inhibited for 3 weeks by 2.5 mM compared to 0.5 mM calcium. The inhibition was 50% on day 3 of culture, and decreased to 19% by day 21. When cultures were incubated with [3H]leucine, radioactive PTH and COOH-terminal PTH fragments were secreted. Sequence analyses were performed on material in radioactive and immunoreactive peaks following gel filtration and high performance liquid chromatography of media. The results indicated that cleavage of PTH or fragments thereof occurred at the 23-24, 27-28, and 33-34 peptide bonds. NH2-terminal fragments of PTH were not detected in media.